tbuckley/cabot-search huggingface dataset:
Abstract P3024: Endothelial To Mesenchymal Transition Frequency Increases In Williams Syndrome Pulmonary Artery Endothelial Cells In Novel 3D Bioprinted <i>In Vitro</i> Arterial Model
Authors: Maher R Saadeh, Martin L Tomov, Lakshmi P Dasi, Reza Avazmohammadi, Vahid Serpooshan, Holly Bauser Heaton
Journal: Circulation Research (2023)

Introduction: Pulmonary artery stenosis (PAS) affects 1 in 2000 live births worldwide, with several causes, most commonly by genetic mutations that lead to or are associated with other congenital heart diseases (CHD). Despite its prevalence, long-term treatment strategies for patients with PAS remains ineffective due to its heterogenous etiology. In the last few decades, researchers have found hemodynamic forces to be a great contributor to stenosis development because it induces endothelial cell (EC) dysfunction through endothelial-to-mesenchymal transition (EndMT). Current pulmonary artery models, in vivo and in vitro , are ineffective in uncovering this mechanism in PAS because they either are unable to recapitulate the phenotype or the micromechanical environment. Methods: To address these limitations, we developed and optimized a high-throughput 3D bioprinted in vitro vascular model that recapitulates patient-specific disease states to determine how EndMT plays a role in stenosis development. This bioprinted model will be a cellularized bilayer comprised of EC and smooth muscle cells (SMC) sourced from either human primary cells or our established Williams Syndrome (WS) induced-pluripotent stem cell (iPSC) lines. WS was chosen due to its high predisposition in developing PAS, around 50%. Our hypothesis is that utilizing our 3D bioprinted model, we will demonstrate how alterations in hemodynamic forces, specifically flow regiments, will further promote EndMT in the WS group. Results: Encapsulated SMC functionality and differentiated phenotype was confirmed by calcium measurement and immunohistochemistry, respectively. Single-cell RNA sequencing (scRNA-seq) was used to examine EC, SMC, EC-SMC crosstalk, and EndMT gene expression profiles. Conclusion: This work represents a new and substantive departure from conventional in vitro vascular modeling, for it will allow us to identify initial vascular cellular responses in acute non-homeostatic conditions and uncover unique signaling pathways found in multiple CHD related PAS. This fundamental knowledge from our vascular mimics can then be applied to facilitate translational advancements in treatment of PAS via surgical and transcatheter intervention methods.
Abstract P3026: Multimodal Imaging To Identify Pre-signals For Acute Aortic Dissection By Using Marfan Syndrome Model Mouse
Authors: KAORI SUGIYAMA, Masahiro Ando, Hiroko Matsunaga, Ryota Wagatsuma, Haruki Uchino, Kenichi Kimura, Katja Schenke-layland, Makoto Arita, Hiromi Yanagisawa, Haruko Takeyama
Journal: Circulation Research (2023)

Background: Marfan syndrome (MFS) is a genetic disorder caused by mutations in the gene encoding theextracellular matrix Fibrillin-1 (FBN1), resulting in systemic connective tissue abnormalities. The mostcommon life-threatening conditions are aortic rupture and/or dissection. Aortic dissections show a highmortality rate, but the details of molecular triggers are unknown, particularly in acute aortic dissection(AAD). Raman spectroscopy is a technique for analyzing molecular structures from Raman spectra. It hasrecently attracted attention in the biology field because it can non-invasively distinguish between diseaseand non-disease areas. Objectives: This study aimed to identify the signals that induce changes in the aortic wall prior to acuteaortic dissection using genetic analysis and Raman spectroscopy with preservation of positional information. Methods: An acute aortic dissection model was created by using MFS model mice (Fbn1 mgR/mgR ) byadministration of angiotensin II (AngII) via osmotic pump (Fbn1 mgR/mgR -AAD). Aortas from wild-type(WT), Fbn1 mgR/mgR and Fbn1 mgR/mgR -AAD mice were harvested, and frozen sections were prepared. Thevascular wall of each mouse was assessed histologically, followed by spatial transcriptomic analysis andRaman microscopy measurements. Multivariate data analysis was performed on the obtained Ramanspectra. Mass spectrometry (MS) imaging was utilized for lipidome analysis. Results: Label-free imaging of extracellular substrates (collagen fibres, elastic fibres, versican, andaggrecan), lipids, and cells was produced in WT, Fbn1 mgR/mgR and Fbn1 mgR/mgR - AAD aortic tissue. Spatialtranscriptomic analysis identified differentially expressed genes among these aortas. We identified markedlipid accumulations in the acute aortic dissection models with Raman and MS imaging.
Abstract P2176: Development Of Integrated Transparent Transgenic Zebrafish Modeling System For Experimental Cardio-oncology Therapeutics
Authors: Surendra K Rajpurohit, Logan Ouellette, Vaani Balyan, Keshu Bhatt, Aiden Van Derhei, Karen Aikhionbare, Raymond Chen, Taitum Gossman, Sai Nasanally, Balakrishna Lokeshwar, Vishal Arora
Journal: Circulation Research (2023)

The zebrafish model has provided an excellent platform to study the genetic and molecular approach of basic and translational cardiac research. Zebrafish heart cells are similar to human heart cells at the molecular level and determine the function of genes that control cardiac function and dysfunction. In zebrafish, myl7 is myosin light chain-7 gene and identified as a regulatory gene of heart orthologs to human MYL7. In vertebrates, including zebrafish, murine and human systems, the in-vivo spatial resolution is limited due to the normal opacification of skin and subdermal structures. For in-vivo imaging the skin transparency is primary requirement and to maintain the transparency, blocking the pigmentation needs to maintain. Chemical inhibition treatment is temporary and possible till the organism treated with the chemical inhibitor agent. Zebrafish casper mutant maintain transparency throughout the life and serve as ideal combination of sensitivity and resolution for in-vivo stem cell analyses and in-vivo imaging. To maintain the skin transparency, in this study, we have developed transparent transgenic zebrafish model and established of time lapse in-vivo confocal microscopy to study of cellular phenotype/pathologies of the cardiomyocytes to quantify changes in cardiomyocyte morphology and function overtime by comparing control and cardiac injury by generating casper/ myl7:RFP; annexin-5:YFP transgenic zebrafish. In the field of cardio-oncology, we are developing in-vivo transparent transgenic cellular phenotype model of zebrafish to study of the cell death process (Annexin5) and Inflammatory activity (NFkB) in the microglia, and the resident macrophage of the brain and cardiomyocytes) homozygous casper/ myl7:RFP; annexin-5:YFP/NF-kB:GFP/mpeg1:mcherryFP transgenic strain zebrafish. By using this novel model our lab establishing the role of NFkB and microglia function under normal healthy status and in glioblastomas Our strategist approach to yield crucial new insights into in-vivo cardiomyocyte imaging by confocal microscopy to observe and track the cell death pattern and cardio inflammatory pathways in cardiomyocyte and develop novel therapeutic approaches to treat cardio inflammatory pathology and glioblastoma
Abstract P3030: Reduced Exercise Capacity Occurs Before Intrinsic Changes In Skeletal Muscle In Experimental Rat Models Of Pulmonary Hypertension
Authors: Peng Zhang, Denielli da Silva Goncalves Bos, Alexander Vang, Julia Feord, Danielle McCullough, Alexsandra Zimmer, Natalie D'Silva, Richard T Clements, Gaurav Choudhary
Journal: Circulation Research (2023)

Background: Pulmonary Hypertension (PH) is defined with mean pulmonary arterial pressure greater than 20 mmHg at rest. Reduced exercise capacity in PH significantly impacts quality of life. While the cause of reduced exercise capacity in PH remains unclear, it is suggested that regional skeletal muscle adaptions may play a role. The objective of this study was to investigate whether intrinsic changes in skeletal muscle determine exercise capacity in PH using preclinical PH rat models with different PH severity. Methods and Results: PH was induced in adult Sprague-Dawley (SD) or Fischer (CDF) rats with one dose of SU5416 (20 mg/kg) injection, followed by 3 weeks of hypoxia and additional 1-4 weeks of normoxia exposure. Rats injected with vehicle and housed in normoxia were used as controls. Echocardiography and hemodynamics were performed to assess PH and cardiac function. Exercise capacity was assessed by VO 2 max. Skeletal muscle structural changes (atrophy, fiber type switch, and capillary density), mitochondrial function, isometric force, and fatigue profile were assessed in soleus muscles. In SD rats, right ventricular systolic dysfunction is associated with reduced exercise capacity in PH rats at 7 weeks timepoint in comparison to control rats, while no changes were observed in skeletal muscle structure, mitochondrial function, isometric force, or fatigue profile. CDF rats at 4 weeks timepoint developed a more severe PH and, in addition to right ventricular dysfunction, the reduced exercise capacity in these rats is associated with skeletal muscle atrophy; however, mitochondrial function, isometric force, and fatigue profile in skeletal muscle remain unchanged. Conclusions: Our study demonstrated that reduced exercise capacity in PH occurs before intrinsic changes in skeletal muscle, suggesting that alterations in skeletal muscle are not causative to exercise intolerance in PH, while they may worsen exercise capacity collaboratively with primary factors in the late stage of PH. In addition, our study supports the notion that improving central cardiopulmonary impairments at the early stage of PH may be critical in preventing or delaying the occurrence of exercise limitation in settings of PH.
Abstract P1085: Generating Injectable Vascularized Cardiomyocytes Derived From Human Induced Pluripotent Stem Cells
Authors: Junya Aoyama, Morgan Sokol, Anthony Zhu, Fang Cao, Eun S Gil, Elton Aleksi, Jessica Garbern, Hannah Elwell, Irving Barrera, Richard T Lee
Journal: Circulation Research (2023)

Introduction: Regenerated cardiomyocytes (CMs) derived from human-induced pluripotent stem cells (hiPS-CMs) have potential for improving deficits in cardiac function due to loss of cardiomyocytes. However, hiPS-CMs are immature relative to adult cardiomyocytes. Here, we demonstrate that injecting CMs with self-assembling peptide (SAP) nanofibers can promote vascularization, organization of sarcomeres, and maturation of the CMs. Methods: We first cultured hiPS-CMs on different SAPs in vitro and analyzed gene expression related to CM maturation. We then implanted hiPS-CMs with the SAP into uninjured rat left ventricular (LV) wall and evaluated sarcomere organization and vascularization up to three months. In addition to uninjured myocardium, we also evaluated the effects of SAPs in post-ischemic rat myocardium. Furthermore, we analyzed the benefit of SAPs on CM maturation of injected hiPS-CMs within the rat LV wall by the Slide-seq spatial single-cell genomics technique. Results: Among SAPs tested, the RADA16 peptide promoted mRNA expression related to CM maturation, including expression of myofibril isoform subunit MYH7/6 , gap junction GJA1 , and potassium ion channel KCNJ2 . Moreover, the expression of HCN4 , a gene expressed in pacemaker cells, decreased on RADA16. The hiPS-CMs injected with RADA16 had improved organization of sarcomeres and more vascularization up to 3 months. More vascularization by RADA16 with hiPS-CMs was also confirmed in the ischemic LV wall. Slide-seq revealed that the maturation of hiPS-CMs occurred earlier with co-injection with the RADA16. Conclusion: The SAP RADA16 promoted improved maturation of CMs in vitro. Furthermore, this SAP promoted the organization of sarcomeres, maturation of myofibrils, and vascularization in vivo, which could benefit implanted hiPS-CMs in improving host cardiac function.
Abstract P1082: The Role Of Shroom3 Protein In Cardiac Regeneration And Repair
Authors: Amirala Bakhshian Nik, Ayana Jamal, Brian Link, Caitlin Omeara
Journal: Circulation Research (2023)

The cardiac injury response mainly depends on species and developmental stage. In a pro-regenerative context, including zebrafish and neonatal mice, following cardiac injury and death of cardiomyocytes (CMs), remaining CMs re-establish a functional cytoarchitecture, proliferate, and integrate into a re-muscularized heart. However, in a non-regenerative context, existing CMs do not readily proliferate, but instead, remodel their cytoarchitecture and undergo hypertrophy. Of note, CM cytoskeletal structure and intercalated disc (ICD) play a key role in regulating cell signaling and cardiac function. In zebrafish, Shroom3 (Shrm3) is a protein that mediates CM morphology, proliferation, and ICD integrity. Previous studies suggest that Shrm3 deletion results in increased CM proliferation in embryonic mice and zebrafish. Here, we investigate the function of Shrm3 on CM and its role in mediating a regenerative response to myocardial injury. To date, the role of Shrm3 in regulating the postnatal CM cell cycle has not been explicitly explored due to high lethality in global Shrm3 knockout mouse models. Thus, we developed a tamoxifen-inducible CM-specific Shrm3 deletion mouse model (Shrm3 fl/fl ;Myh6 MCM ) to study the Shrm3 role in both neonate and adult mice. At P7, Shrm3 fl/fl ;Myh6 MCM uninjured mice showed increased EdU incorporation in CM nuclei compared to wildtype controls suggesting that as in zebrafish and embryonic mice, Shrm3 knockout increased CM cell cycle activity postnatally. We performed apical resection injury on P1 mice and at 21 days post-injury (dpi) Shrm3 fl/fl ;Myh6 MCM CMs displayed compromised ICDs. Trichrome staining of 21 dpi hearts revealed minimal scar formation in Shrm3 fl/fl ;Myh6 MCM mice, however, we observed aberrant regeneration of the ventricular apex as indicated by invagination of the cardiac apex and significantly reduced ejection fraction at 21 dpi. Thus, our data suggest that Shrm3 deletion in postnatal CMs remodels cell junctions and increases CM cell cycle activity. Our future studies will address the Shrm3 function(s) in cardiac regeneration and repair in injured adult mice to develop potential therapeutic strategies.
Abstract P3144: A Deep Neural Network Algorithm For Accurate Detection Of Brugada Syndrome Features In Electrocardiograms
Authors: Luke Melo, Giuseppe Ciconte, Ashton Christy, Gabriele Vicedomini, Luigi Anastasia, Edward Grant, Carlo Pappone
Journal: Circulation Research (2023)

Brugada Syndrome (BrS) is a rare but potentially fatal cardiac condition that is difficult to diagnose due to the elusive nature of its characteristic ECG pattern. The diagnosis of BrS requires identification of a specific electrocardiographic (ECG) pattern, which often appears only upon administration of sodium-channel blockers. Here, we present a deep neural network (DNN) algorithm that accurately detects BrS features in ECGs without the need for a drug challenge. We trained and validated the DNN on two independent cohorts of patients who were enrolled in electrophysiological studies for BrS diagnosis. The internal prospective validation cohort included 370 subjects, and the multicenter external validation cohort included 110 subjects from three other medical institutions in Italy. All subjects who did not present a spontaneous type 1 ECG underwent an ajmaline drug challenge to aid in confirming a BrS diagnosis. We found that the DNN approach classified ECGs for BrS with a sensitivity of 79.6%, specificity of 93.6%, accuracy of 88.4%, and area under the curve (AUC) of 0.934±0.027 in the validation cohorts. The DNN model also detected BrS with 100% accuracy in all cases in which a patient presented the spontaneous type 1 pattern. Our results demonstrate that a multivariate machine-learning algorithm can accurately detect the features of BrS in a conventional ECG without the challenge of a sodium-channel blocker. The computer-assisted analysis of digital ECG traces from current electrocardiographs might help physicians recognize patients affected by this disease. In conclusion, we have developed an advanced deep learning algorithm that can accurately detect BrS features in ECGs. Our results demonstrate that the DNN approach could be an effective tool to improve the diagnosis of BrS and potentially prevent sudden cardiac death in affected individuals.
MST4 kinase regulates immune thrombocytopenia by phosphorylating STAT1-mediated M1 polarization of macrophages
Authors: Jingjing Cao, Lili Ji, Yanxia Zhan, Xia Shao, Pengcheng Xu, Boting Wu, Pu Chen, Luya Cheng, Xibing Zhuang, Yang Ou, Fanli Hua, Lihua Sun, Feng Li, Hao Chen, Zhaocai Zhou, Yunfeng Cheng
Journal: Cellular and Molecular Immunology (2023)
What are the chances of having a second epileptic seizure?
Authors: Helen Saul, Brendan Deeney, Samantha Cassidy, Aidan Neligan
Journal: BMJ (2023)

The studyNeligan A, Adan G, Nevitt SJ, et al. Prognosis of adults and children following a first unprovoked seizure. Cochrane Database Syst Rev 2023;1:1-75.To read the full NIHR Alert, go to: https://evidence.nihr.ac.uk/alert/epilepsy-what-are-the-chances-of-having-a-second-seizure/.
Subclinical Keratoconus Detection and Characterization Using Motion-Tracking Brillouin Microscopy
Authors: J. Bradley Randleman, Hongyuan Zhang, Lara Asroui, Imane Tarib, William J. Dupps, Giuliano Scarcelli
Journal: Ophthalmology (2023)

PurposeTo characterize focal biomechanical alterations in subclinical keratoconus (SKC) using motion tracking (MT) Brillouin microscopy and evaluate the ability of MT Brillouin metrics to differentiate SKC eyes from normal controls.DesignProspective cross-sectional studyParticipantsThirty eyes from 30 patients were evaluated, including 15 eyes from 15 bilaterally normal patients and 15 SKC eyes from 15 patients.MethodsAll patients underwent Scheimpflug tomography and MT Brillouin microscopy imaging using a custom-built device. Mean and Minimum (Min) MT Brillouin values within the anterior plateau region (Plateau) and anterior 150 microns (A150) were generated. Scheimpflug metrics evaluated included Inferior-Superior (IS) Value, K Max, Thinnest Corneal Thickness (TCT), asymmetry indices, BAD-D, and ART max. Receiver operating characteristic (ROC) curves were generated for all Scheimpflug and MT Brillouin metrics evaluated to determine the area under the curve (AUROC), sensitivity, and specificity for each variable.Main Outcome MeasuresDiscriminative performance based on AUROC, sensitivity, and specificity.ResultsThere were no significant differences between groups for age, sex, manifest refraction spherical equivalent refractions (MRSE), corrected distance visual acuity CDVA, K Max, or KISA% index. Among Scheimpflug metrics, there were significant differences between groups for TCT (556μm vs. 522μm, p < 0.001), IS Value (0.29D v s 1.05D, p <0.001), IVA (0.10 vs 0.19, p <0.001) and KI (1.01 vs 1.05, p <0.001), and no significant differences for any other Scheimpflug metric. Among MT Brillouin metrics, there were clear differences between Control and SKC eyes for Mean Plateau (5.71 GHz vs. 5.68 GHz, p<0.0001), Min Plateau (5.69 GHz vs. 5.65 GHz, p<0.0001), Mean A150 (5.72 GHz vs. 5.68 GHz, p<0.0001) and Min A150 (5.70 GHz vs. 5.65 GHz, p<0.001). All MT Brillouin Plateau and A150 Mean and Min metrics fully differentiated groups (AUROC = 1.0 for each), while the best performing Scheimpflug metrics were the Keratoconus Index (AUROC = 0.91), IS Value (AUROC = 0.89), and Index of Vertical Asymmetry (IVA, AUROC = 0.88).ConclusionsMT Brillouin microscopy metrics effectively characterized focal corneal biomechanical alterations in SKC eyes and clearly differentiated these eyes from controls, including eyes that were not accurately differentiated using Scheimpflug metrics.
Abstract P3195: Mitochondrial CaMKII Drives Cardiometabolic Dysfunction In Heart Failure
Authors: Kimberly Ferrero, Robert Cole, D B Foster, Jonathan Granger, Brian O'Rourke, David A Kass, Elizabeth Luczak
Journal: Circulation Research (2023)

Background: Ca 2+ /calmodulin-dependent protein kinase II (CaMKII) is an established negative regulator of cardiac injury. Both the expression and activity levels of CaMKII delta, the primary cardiac isoform of this kinase, are elevated in models of heart failure (HF) such as ischemia-reperfusion (I/R) injury and following myocardial infarction (MI). This upregulation is due in part to CaMKII’s role in the regulation of excitation-contraction coupling, apoptosis, activation of hypertrophic programming, arrhythmias and pro-inflammatory signaling. Recently, a pool of mitochondrial CaMKII (mtCaMKII) has been identified; we have demonstrated that there is an observed increase in mitochondrial CaMKII activation with left ventricular dilation acutely following injury in a mouse model of MI. This deleterious post-MI phenotype is rescued with genetic mitochondrial CaMKII inhibition; conversely, mice with myocardial mitochondrial CaMKII overexpression present with a severe dilated cardiomyopathy phenotype, alterations in electron transport chain (ETC) activity, and decreased ATP production. Objective: To date, the molecular mechanisms by which CaMKII regulates mitochondrial energetics are unknown and present a novel therapeutic target for HF which we are currently studying. Methods: As we have identified changes in the activity of enzymes, particularly complexes I and II, in the mitochondrial ETC as well as TCA cycle in response to increased CaMKII levels or activity, we are in the process of assessing changes in the mitochondrial CaMKII interactome between uninjured and failing hearts. Utilizing liquid chromatography-mass spectrometry (LCMS) and proteomics analysis of both scaffolding interactions alongside a novel ATP analogue labeling technique, we have mapped novel mitochondrial proteins which interact with mtCaMKII. Significance: The identification of previously unknown mitochondrial pathways involving CaMKII may uncover promising pharmacological targets for cardiovascular therapeutics, as this kinase is a critical player in the progression of HF.